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Methods and Assays for Specific Targeting and Delivery of RNA Nanoparticles to Cancer Metastases

  • Piotr Rychahou
  • Yi Shu
  • Farzin Haque
  • Jiyao Hu
  • Peixuan Guo
  • B. Mark Evers
Part of the Methods in Molecular Biology book series (MIMB, volume 1297)

Abstract

In recent years, RNA nanotechnology has become increasingly attractive due to its potential for applications in nanomedicine. RNA nanotechnology refers to the design and synthesis of nanoparticles composed mainly of RNA via bottom-up self-assembly. RNA nanoparticle is a suitable candidate for targeted delivery of therapeutics to cancer cells due to its multivalency, which allows the combination of therapeutic, targeting, and detection moieties all into one nanoparticle. To date, a system capable of exclusively targeting metastatic cancers that have spread to distant organs or lymph nodes is in demand. In this chapter, we report methods for establishing the clinically relevant colorectal cancer mouse metastasis models and describe methods and assays for constructing multifunctional, thermodynamically and chemically stable RNA nanoparticles that specifically target colorectal cancer metastases in the liver. Systemic injection of RNA nanoparticles showed metastatic cells targeting with little or no accumulation in normal liver parenchyma several hours after injection, demonstrating the therapeutic potential of these RNA nanoparticles as a delivery system for the treatment of cancer metastases.

Key words

RNA nanotechnology RNA nanoparticles Cancer metastasis RNA therapeutics Specific delivery 

Notes

Acknowledgements

The research was supported by NIH grants R01 DK048498, P30 CA177558, and The Markey Cancer Foundation to B.M.E, as well as R01 EB003730 and U01 CA151648 to P.G. The content is solely the responsibility of the authors and does not necessarily represent the official views of NIH. Funding to Peixuan Guo’s Endowed Chair in Nanobiotechnology position is from the William Fairish Endowment Fund. P.G. is a cofounder of Kylin Therapeutics, Inc., RNA Nano, LLC., and Biomotor and Nucleic Acid Nanotechnology Development Corp., Ltd.

References

  1. 1.
    Guo P (2010) The emerging field of RNA nanotechnology. Nat Nanotechnol 5:833–842CrossRefGoogle Scholar
  2. 2.
    Guo P, Haque F, Hallahan B et al (2012) Uniqueness, advantages, challenges, solutions, and perspectives in therapeutics applying RNA nanotechnology. Nucleic Acid Ther 22:226–245Google Scholar
  3. 3.
    Shu Y, Pi F, Sharma A et al (2014) Stable RNA nanoparticles as potential new generation drugs for cancer therapy. Adv Drug Deliv Rev 66C:74–89CrossRefGoogle Scholar
  4. 4.
    Guo P, Haque F (eds) (2013) RNA Nanotechnology and Therapeutics. Press, CRCGoogle Scholar
  5. 5.
    Guo P, Zhang C, Chen C et al (1998) Inter-RNA interaction of phage phi29 pRNA to form a hexameric complex for viral DNA transportation. Mol Cell 2:149–155CrossRefGoogle Scholar
  6. 6.
    Shukla GC, Haque F, Tor Y et al (2011) A Boost for the Emerging Field of RNA Nanotechnology. ACS Nano 5:3405–3418CrossRefGoogle Scholar
  7. 7.
    Leontis N, Sweeney B, Haque F et al (2013) Conference Scene: Advances in RNA nanotechnology promise to transform medicine. Nanomedicine 8:1051–1054CrossRefGoogle Scholar
  8. 8.
    Kruger K, Grabowski PJ, Zaug AJ et al (1982) Self-splicing RNA: autoexcision and autocyclization of the ribosomal RNA intervening sequence of Tetrahymena. Cell 31:147–157CrossRefGoogle Scholar
  9. 9.
    Guerrier-Takada C, Gardiner K, Marsh T et al (1983) The RNA moiety of ribonuclease P is the catalytic subunit of the enzyme. Cell 35:849–857CrossRefGoogle Scholar
  10. 10.
    Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553CrossRefGoogle Scholar
  11. 11.
    Carmichael GG (2002) Medicine: silencing viruses with RNA. Nature 418:379–380CrossRefGoogle Scholar
  12. 12.
    Winkler WC, Nahvi A, Roth A et al (2004) Control of gene expression by a natural metabolite-responsive ribozyme. Nature 428:281–286CrossRefGoogle Scholar
  13. 13.
    Mulhbacher J, St-Pierre P, Lafontaine DA (2010) Therapeutic applications of ribozymes and riboswitches. Curr Opin Pharmacol 10:551–556CrossRefGoogle Scholar
  14. 14.
    Chen Y, Zhu X, Zhang X et al (2010) Nanoparticles modified with tumor-targeting scFv deliver siRNA and miRNA for cancer therapy. Mol Ther 18:1650–1656CrossRefGoogle Scholar
  15. 15.
    Pegtel DM, Cosmopoulos K, Thorley-Lawson DA et al (2010) Functional delivery of viral miRNAs via exosomes. Proc Natl Acad Sci U S A 107:6328–6333CrossRefGoogle Scholar
  16. 16.
    Fire A, Xu S, Montgomery MK et al (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811CrossRefGoogle Scholar
  17. 17.
    Aagaard L, Rossi JJ (2007) RNAi therapeutics: Principles, prospects and challenges. Adv Drug Deliv Rev 59:75–86CrossRefGoogle Scholar
  18. 18.
    Cerchia L, de Franciscis V (2010) Targeting cancer cells with nucleic acid aptamers. Trends Biotechnol 28:517–525CrossRefGoogle Scholar
  19. 19.
    Keefe AD, Pai S, Ellington A (2010) Aptamers as therapeutics. Nat Rev Drug Discov 9:537–550CrossRefGoogle Scholar
  20. 20.
    Shu D, Shu Y, Haque F et al (2011) Thermodynamically stable RNA three-way junctions for constructing multifuntional nanoparticles for delivery of therapeutics. Nat Nanotechnol 6:658–667CrossRefGoogle Scholar
  21. 21.
    Haque F, Shu D, Shu Y et al (2012) Ultrastable synergistic tetravalent RNA nanoparticles for targeting to cancers. Nano Today 7:245–257CrossRefGoogle Scholar
  22. 22.
    Shu Y, Haque F, Shu D et al (2013) Fabrication of 14 Different RNA Nanoparticles for Specific Tumor Targeting without Accumulation in Normal Organs. RNA 19:766–777CrossRefGoogle Scholar
  23. 23.
    Shu Y, Shu D, Haque F et al (2013) Fabrication of pRNA nanoparticles to deliver therapeutic RNAs and bioactive compounds into tumor cells. Nat Protoc 8:1635–1659CrossRefGoogle Scholar
  24. 24.
    Liu J, Guo S, Cinier M et al (2010) Fabrication of stable and RNase-resistant RNA nanoparticles active in gearing the nanomotors for viral DNA packaging. ACS Nano 5:237–246CrossRefGoogle Scholar
  25. 25.
    Abdelmawla S, Guo S, Zhang L et al (2011) Pharmacological characterization of chemically synthesized monomeric pRNA nanoparticles for systemic delivery. Mol Ther 19:1312–1322CrossRefGoogle Scholar
  26. 26.
    Guo S, Huang F, Guo P (2006) Construction of folate-conjugated pRNA of bacteriophage phi29 DNA packaging motor for delivery of chimeric siRNA to nasopharyngeal carcinoma cells. Gene Ther 13:814–820CrossRefGoogle Scholar
  27. 27.
    Guo S, Tschammer N, Mohammed S et al (2005) Specific delivery of therapeutic RNAs to cancer cells via the dimerization mechanism of phi29 motor pRNA. Hum Gene Ther 16:1097–1109CrossRefGoogle Scholar
  28. 28.
    Shu D, Moll WD, Deng Z et al (2004) Bottom-up assembly of RNA arrays and superstructures as potential parts in nanotechnology. Nano Lett 4:1717–1723CrossRefGoogle Scholar
  29. 29.
    Khisamutdinov EF, Jasinski DL, Guo P (2014) RNA as a boiling-resistant anionic polymer material to build robust structures with defined shape and stoichiometry. ACS Nano 8:4771–4781CrossRefGoogle Scholar
  30. 30.
    Siegel R, Ma J, Zou Z et al (2014) Cancer statistics, 2014. CA Cancer J Clin 64:9–29CrossRefGoogle Scholar
  31. 31.
    Siegel R, Desantis C, Jemal A (2014) Colorectal cancer statistics, 2014. CA Cancer J Clin 64:104–117CrossRefGoogle Scholar
  32. 32.
    Wanebo HJ, Semoglou C, Attiyeh F et al (1978) Surgical management of patients with primary operable colorectal cancer and synchronous liver metastases. Am J Surg 135:81–85CrossRefGoogle Scholar
  33. 33.
    Yoon SS, Tanabe KK (1999) Surgical treatment and other regional treatments for colorectal cancer liver metastases. Oncologist 4:197–208Google Scholar
  34. 34.
    Rychahou P, Haque F, Shu Y et al (2015) Delivery of RNA nanoparticles into colorectal cancer metastases following systemic administration. ACS Nano 9:1108–1116Google Scholar
  35. 35.
    van Dam PA, Watson JV, Lowe DG et al (1990) Tissue preparation for simultaneous flow cytometric quantitation of tumour associated antigens and DNA in solid tumours. J Clin Pathol 43:833–839CrossRefGoogle Scholar
  36. 36.
    Ferreira-Facio CS, Milito C, Botafogo V et al (2013) Contribution of multiparameter flow cytometry immunophenotyping to the diagnostic screening and classification of pediatric cancer. PLoS One 8:e55534CrossRefGoogle Scholar
  37. 37.
    Sukhdeo K, Paramban RI, Vidal JG et al (2013) Multiplex flow cytometry barcoding and antibody arrays identify surface antigen profiles of primary and metastatic colon cancer cell lines. PLoS One 8:e53015CrossRefGoogle Scholar
  38. 38.
    Ricci-Vitiani L, Lombardi DG, Pilozzi E et al (2007) Identification and expansion of human colon-cancer-initiating cells. Nature 445:111–115CrossRefGoogle Scholar
  39. 39.
    Karlsson M, Nilsson O, Thorn M et al (2008) Detection of metastatic colon cancer cells in sentinel nodes by flow cytometry. J Immunol Methods 334:122–133CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Piotr Rychahou
    • 1
  • Yi Shu
    • 2
  • Farzin Haque
    • 3
  • Jiyao Hu
    • 4
  • Peixuan Guo
    • 5
  • B. Mark Evers
    • 6
  1. 1.Markey Cancer Center, Department of SurgeryUniversity of KentuckyLexingtonUSA
  2. 2.Nanobiotechnology Center, Department of Pharmaceutical SciencesUniversity of KentuckyLexingtonUSA
  3. 3.Nanobiotechnology Center, Markey Cancer Center, Department of Pharmaceutical SciencesUniversity of KentuckyLexingtonUSA
  4. 4.Integrated Oncology Laboratory Corporation of AmericaPhoenixUSA
  5. 5.Nanobiotechnology Center, Markey Cancer Center, Department of Pharmaceutical SciencesUniversity of KentuckyLexingtonUSA
  6. 6.Markey Cancer CenterUniversity of KentuckyLexingtonUSA

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